Tuesday, November 21, 2006

BoZZ MukS mAh TheSis

___________________A Thesis Presented toThe Faculty of the College of Arts and SciencesNatural Science DepartmentAteneo de Zamboanga UniversityZamboanaga City___________________In Partial Fulfilmentof the Requirement for theDegree of Bachelor of Science in Biology

By:Hadjula, Mukramel M.

Chapter IIntroduction

Rationale of the study

The uses of traditional medicinal plants in relieving symptoms of disease and curing various infections date back many centuries. In recent years, considerable interest has developed in Asian countries in the collection and extended use of indigenous and introduced Plants for medicinal purposes. Knowledge of medicinal plants has been the key for the survival of the ethnic groups who live in the interior. Peoples who live far from towns and in forest still rely on traditional cures handed down to them through the generations. They use these plants in one form or another to cure or alleviate a variety of ills, for example, toothache, stomach-ache and cough.The control of gastrointestinal nematodes in ruminants has long been relying on the repeated prophylactic use of broad spectrum anthelmintics (AH), sometimes completed with some grazing management. However, the constant development and diffusion of AH resistance within worm populations is now seriously questioning the sustainability of this mode of chemical control. The phenomenon of AH resistance is worldwide and has been described both in temperate and tropical areas (Jackson and Coop, 2000). Therefore, the need to develop alternative methods to AHs has been underlined since several years (Waller, 1999).For tropical countries, a promising alternative to conventional chemotherapy, a promising approach is represented by the scientific evaluation of plants which are traditionally use against digestive troubles and or helminths. Several recent reviews have concerned the ethno veterinary medicine from several continents (Hammond et al, 1997; Sidi Ba 1994; Alawa et al, 2003).The anthelmintic alternative plants are not commonly use in zamboanga. Anthelmintic discovery is both beneficial to humans and animals. Makahiya is known as a diuretic, and is considered alterant and antiasthmatic. It is used for urinary complaints, and is useful in diseases arising from corrupt blood and bile. This finding, stimulate the researcher to attempt to study the anthelmintic effect of Makahiya (Mimosa pudica) leaves extract in Native Chicken (Gallus domesticus) naturally infected with Gastro-intestinal Parasites.

Statement of the problem

The study aims to determine the anthelminthic effect of makahiya (Mimosa pudica) leaves extract, specifically it answer the following question,

Highly educated people living in urban cities and surrounded by modernized hospitals seemed to ignore the importance of herbal medicinal plants like Makahiya (Mimosa pudica), more so those who are well of do not subscribe to any medicinal plants used by ethnic groups.However due to increase and high prices of medicines sold at the drugstores people who belong to the poverty status are compelled to use medicinal plants.Relevant to the uses of medicinal plants, the result of this study will inform the people especially the farmers and poultry raisers. It is both beneficial to humans and animals. Makahiya will become as an alternative anthelmintic in humans and animals like in poultry raising. It is readily available in our surrounding and can be prepared easily. It will reduce our budget in poultry raising and in our daily expenses. The study is served as the first step in the discovery of the bioactive compounds of Makahiya leaves. Finally, these serve as references for other researchers of the similar study.

Scope and limitation

The area of this study is limited to the anthelmintic effect of Makahiya leaves Extract in Native Chicken (Gallus domesticus) naturally infected with Gastro-intestinal Parasites. It focuses on the fecal analysis of sample chicken using Mc Master Egg counting. Only 50 sample chicken used in the experiment. The collection of the feces sample is up to 5 weeks only.

Chapter II

Review of related literature

Native chickens are still very common in the backyards of most rural people in the Philippines.The native chicken has evolved in a way that allows it to survive and reproduce in a marginal environment and with minimal management. More important, the meat of the native chicken has a unique flavor and texture which consumers prefer, and pay a premium price for. To date, the native chicken remains an important source of high-quality protein food and additional income for many rural dwellers. Furthermore, it performs other socio-economic and cultural roles i.e. as a form of savings and insurance, as a buffer against periodic shortages, and as a way of diversifying farm resources and allowing low-income farmers to meet their social and cultural obligations.Concern over food safety and health issues has resulted in a shift in consumer preferences towards meat products with good flavor, from chickens raised in a more humane environment, with minimal antibiotics or chemical-based feed additives. These new preferences have opened opportunities for financially challenged rural farmers to engage in native chicken production. However, wide variation in the performance of native chickens and the quality of their products is a constraint to its utilization on a larger scale.Several technologies have been tested and shown to improve the productivity of native chickens. However, most of these require financial and technical inputs that are far beyond the capacity of poor farmers. Nonetheless, some technologies that have been developed by small-scale farmers themselves have been shown to improve productivity and product quality of native chickens, and at the same time allow farmers to synchronize production so as to meet market demand. (http://www.agnet.org/library/article/pt2003007.html)Ascaridia galli is a nematodeparasite that causes ascariasis, or worm infection, in poultry. Life cycle Eggs have thick, albuminous shell that are resistant to desiccation and persist for a long time in the environment. Larvae do not hatch but moult inside the eggs until it reaches the L3 stage. This can take about two weeks but the period depends on other factors such as weather condition. The infective eggs are ingested by a chicken where it reaches the small intestine and hatches. Temperature, carbon dioxide levels and pH are thought to be triggering factors that signal the larvae to hatch from its egg. The larvae then burrows into the mucosal lining of the small intestine where it undergoes two additional moults. It is this phase of their lifecycle where these worms causes the most damage to their host. They then re-enter the small intestine and develop into adults where they live their lives out feeding on gut content and making a vast amount of eggs that would then be excreted by a host and free to continue their lifecycle. If the animal is able to mount an immune respond to the larvae, i.e. from pre-exposure, the larvae do not develop into adults but hides in the mucosa of the small intestine. This is common for infection of older birds. Transport host such as earthworm is thought to play a role in transmission of Ascaridia galli and hence, free range birds tend to have a higher risk of infection.(http://en.wikipedia.org/wiki/Ascaridia_galli)Chenopodium ambrosioides (Family Chenopodiaceas) originated in Central America, though it has been distributed to much of the world. It has been used as an anthelmintic (medicine for controlling internal parasites) for many years. In the early 1900s it was one of the major anthelmintics used to treat ascarids and hookworms in humans, cats, dogs, horses, and pigs. Usually, oil of chenopodium was used. It was sometimes referred to as Baltimore Oil, because of the large production facility in Baltimore that specialized in extracting the oil from the plant. Chenopodium was replaced with other, more effective and less toxic anthelmintics in the 1940s. Chenopodium is still used to treat worm infections in humans in many countries. In Honduras, as well as other Latin American countries, the whole plant or the leaves are ground and added to water. This mixture is then consumed. In a few areas in Latin America, the plant also is used to treat worm infections in livestock.(http://www.abc.cornell.edu/plants/medicinal/epazote.html)Taken internally, Aloe Vera demonstrates powerful detoxifying effects by flushing out toxins and promoting healthy tissue formation. Traditionally, it has been used in Indian medicine for asthma, jaundice, as a carminative, for various musculoskeletal disorders, for suppression of menstrual disorders, as a tonic, purgative, aphrodisiac, antihelminthic, in various opthalmological disorders, enlargement of the spleen, various forms of hepatitis, vomiting, fever due to bronchitis, and for erysipelas. (http://www.luresext.edu/international/parasite.htm)Allium sativum (garlic) - In vitro, garlic reportedly has nematocidal effects against Ascaris lumbricoides. The active component in garlic is allicin, which appears to interfere with sulfhydryl-dependent enzymes in microorganisms. However, because allicin can be degraded by heat, only large quantities of fresh raw garlic cloves may be useful for the treatment of worm infestations. Although garlic has not yet been subjected to clinical trials in roundworm infections, it is considered to be a safe herb (Heron and Yarnell 1999).Ginger was used historically in different regions of the world for the same basic therapeutic applications. These include: analgesic, anti-arthritic, wound healing, anthelmintic, anti-ulcer, stimulant and aphrodisiac properties, plus treatment of a variety of respiratory, reproductive and digestive complaints. (http://www.herbs2000.com/herbs/herbs_ginger.htm) Stem bark of MANGIFERA INDICA L. Mango has been found to possess antihelminthic and antiallergic properties (Garcia et al., 2003)cassia alata has been used for soothing inflammation and shingles, as a skin disinfectant, for constipation, oedema, herpes infections, hepatitis, liver discomfort, impetigo, worm infestation, as a laxative and an analgesic. Also it has been used for leprosy, wound healing, as an anti-helminthic, anti-bacterial, diuretic, for snakebites, bronchitis, asthma, hasten child birth, eczema, stomachaches and uterine disorders.(http://content.nhiondemand.com/moh/media/monoHerb.asp?objID=101061&ctype=herb&mtyp=1#fn115048)The application of papaya latex that is probably of most interest to livestock producers is as an anthelmintic (dewormer). Satrija et al. (1994) tested the efficacy of papaya latex (at doses of 2, 4, and 8 g/kg BW) against Ascaris suum in 16 pigs. The eggs per gram (epg) on days 0, 1, 5, and 7 were determined using a modified McMaster technique and the adult worms were collected and sexed at necropsy on day 7. The 4- and 8-g/kg BW treatments significantly decreased the epg produced (by 99%) and the number of adult worms by 80 and 100%, respectively. The study conducted by Satrija et al. supports the results of other studies which indicate that papaya latex is effective against Ascaridia galli in chickens. One adverse effect of the treatment was transient diarrhea in the 8-g/kg BW group on day 1 of the study. In another study, water extracts of papaya seeds decreased Ascaridia galli infections in chicks by 41.7% (compared to piperazine hexahydrate which decreased infections by 99%).In traditional veterinary medicine, papaya seeds also are used as dewormers. In Indonesia and the Philippines, air-dried seeds are ground and mixed with water - 3 g of seeds/kg bodyweight. The animals are given the seed/water mixture once a day for 6 days. In Indonesia, papaya leaves are used as affed for animals after parturition - 2 leaves boiled in water fed every 2 days for 1 week. It also has been reported that papaya leaf extract is used as a profilaxis against malaria, though no studies on this use could be found in the literature. (http://www.ansci.cornell.edu/plants/medicinal/papaya.html)The Sensitive plant (Mimosa pudica L.) is a creeping annual or perennial herb often grown for its curiosity value: the compound leaves fold inward and droop when touched, re-opening within minutes. Mimosa pudica is native to Brazil, but is now a pantropical weed. Other names given to this curious plant are Humble plant, Shame plant, Sleeping grass, Prayer Plant, Touch-me-not, Makahiya (Philippines, meaning "shy"), and Mori Vivi (West Indies). The Chinese name for this plant translates to "shyness grass". The species epithet, pudica, is Latin for "bashful" or "shrinking".The stem is erect in young plants, but becomes creeping or trailing with age. The stem is slender, branching, and sparsely to densely prickly, growing to a length of 1.5 m (5 ft). The leaves are bipinnately compound, with one or two pinnae pairs, and 10-26 leaflets per pinna. The petioles are also prickly. Pedunculate (stalked) pale pink or purple flower heads arise from the leaf axils. The globose to ovoid heads are 8-10 mm in diameter (excluding the stamens). On close examination, it is seen that the floret petals are red in their upper part and the filaments are pink to lavender. The fruit consists of clusters of 2-8 pods from 1-2 cm long each, these prickly on the margins. The pods break into 2-5 segments and contain pale brown seeds some 2.5 mm long.Mimosa pudica is well known for its rapid plant movement. In the evening the leaflets will fold together and the whole leaf droops downward. It then re-opens at sunrise. This type of motion has been termed nyctinastic movement. The leaves also close up under various other stimuli, such as touching, warming, or shaking. The stimulus can also be transmitted to neighbouring leaves. These types of movements have been termed seismonastic movements. The cause is a loss of turgor pressure. The movement is caused by "a rapid loss of pressure in strategically situated cells that cause the leaves to droop right before one’s eyes".(http://www.informat.io/?title=sensitive-plant)Makahiya is found throughout the Philippines and is found wild everywhere on Mt. Banahaw. It is not cultivated, and is considered a weed. The Tagalog name, and suggested by the scientific name, means "sensitive." The reason it is given this name is that upon human touch the leaves contract and appear to wilt, but then open up again a short time later. Makahiya is known as a diuretic, and is considered alterant and antiasthmatic. It is used for urinary complaints, and is useful in diseases arising from corrupt blood and bile.(http://www.filipinoherbshealingwonders.filipinovegetarianrecipe.com/herbs_pics/makahiya.htm)Many Filipinos have been patronizing herbal medicines for the past decade or so, and folk medicines for centuries, from the cure of simple cough and cold ("lagundi") to hypertension ("pito-pito"), hyper acidity ("makahiya") and erectile dysfunction ("burong sibuyas na pula"). (Cinco, 2006)In the study conducted in zamboanga city, Comparative Study On The Efficacy Of Ipil-Ipil Seeds Versus Mebendazole In The Treatment Of Intestinal Ascariasis And Trichuriasis Among Elementary Students (Yu, 2004), The antihelminthic effect of Ipil-Ipil seeds was found to be comparable with Mebendazole based on ascaris and trichuris egg reduction rates.

Chapter III

METHODOLOGY

Materials and method

The poultry raising in Sta.Maria is the place where the feces sample is collected. Initial fecalysis were done to ensure if the 50 selected sample chickens is affected with the roundworms. The infected chickens were place into 3 separate groups of cages and labelled. The feces were collected every 3 days for 5 weeks in a clean film container.

Preparation of extract leaves of makahiya

The fresh leaves of makahiya were collected from Ayala and bring to Ateneo research laboratory. The leaves were weigh of about 500g. Makahiya leaves were wash thoroughly using sterile hand gloves and place in mortal. The pulverized leaves were place in a clean cheese cloth and extracted to beaker. The extracted makahiya leaves were diluted in 3 separated 250ml beaker. The beakers were labelled 75%, 50%, and 25% concentration of makahiya leaves. First beaker - contain 25ml of water and a total of 75ml of makahiya leaves represents 70% concentration. Second beaker - contain 50ml of water and a total of 50ml of makahiya leaves represents 50% concentration. Third beaker - contain 75ml of water and a total of 25ml of makahiya leaves represents 25% concentration.

Administration of the treatment

The 50 samples of chicken were divided into 3 groups. First group composed of 30 chickens which dived into three groups composed of 10 chickens to be treated orally using 3ml syringe with 75%, 50% and 25% concentration of extracted makahiya leaves. Second group composed of 10 chicken and untreated. The third group (control) is composed of 10 chicken to be treated orally with commercial anthelminthic (piperazine dihydrochloride) as the control.

Preparation of samples (feces)

The feces were collected every 3 days for 5 weeks from 3 groups of chicken. The collected feces were place in a film clean container and bring to the Department of agriculture in Tumaga. The egg worm count was done using Mc Master Egg counting technique.

Mc Master Egg counting technique

Parasitological examination was done by direct smear, sedimentation, and flotation techniques following the standard procedures. Those fecal samples were subjected to egg output (eggs per gram; EPG) of feces count using Mc Master Egg counting technique at Department of Agriculture in Tumaga.